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Alexa Fluor®647 Mouse Anti-Human CD4
Alexa Fluor®647 Mouse Anti-Human CD4

Multiparameter flow cytometric analysis of CD4 expression on human peripheral blood leucocytes populations. Whole blood was stained with either Alexa Fluor™ 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing CD4 expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Alexa Fluor®647 Mouse Anti-Human CD4

Profile of peripheral blood lymphocytes analyzed by flow cytometry.

Multiparameter flow cytometric analysis of CD4 expression on human peripheral blood leucocytes populations. Whole blood was stained with either Alexa Fluor™ 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing CD4 expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Profile of peripheral blood lymphocytes analyzed by flow cytometry.

Product Details
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BD Pharmingen™
L3T4; T-cell surface antigen T4/Leu-3; W3/25; CD4 antigen (p55)
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
IV T114; VI 6T-079
920
AB_396816
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. An isotype control should be used at the same concentration as the antibody of interest.
557707 Rev. 5
Antibody Details
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RPA-T4

The RPA-T4 monoclonal antibody specifically binds to CD4, a 59 kDa single-chain transmembrane glycoprotein that is expressed on T-helper/inducer cell populations. CD4 is also expressed on thymocyte subsets and at lower levels on monocytes and macrophages. CD4 functions as a co-receptor in MHC class II-restricted antigen-induced T cell activation and as a receptor for human immunodeficiency viruses (HIV). This antibody binds to the D1 domain (CDR1 and CDR3 epitopes) of the CD4 antigen and reacts with approximately 80% of thymocytes and 45% of peripheral blood lymphocytes. RPA-T4 is capable of blocking HIV-1, gp120, and inhibits syncytium formation.

557707 Rev. 5
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
557707 Rev.5
Citations & References
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Development References (3)

  1. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  2. Piatier-Tonneau D. CD4 workshop panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:49-54.
  3. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
557707 Rev. 5

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.